湖南柑橘溃疡病菌的多态性鉴定及致病力分析  

Polymorphism and pathogenicity analysis of citrus canker pathogens in Hunan

作　　者：罗旭钊 朱韬 郝晨星 朱亦赤 马先锋 邓子牛[1,2] 韩健 LUO Xuzhao;ZHU Tao;HAO Chenxing;ZHU Yichi;MA Xianfeng;DENG Ziniu;HAN Jian(Engineering Research Center for Horticultural Crop Germplasm Creation and New Variety Breeding,Changsha,Hunan 410128,China;National Center for Citrus Improvement,Changsha,Hunan 410128,China;Hunan Horticultural Research Institute,Changsha,Hunan 410125,China)

机构地区：[1]园艺作物种质创新与新品种选育教育部工程中心,湖南长沙410128 [2]国家柑橘改良中心长沙分中心,湖南长沙410128 [3]湖南省园艺研究所,湖南长沙410125

出　　处：《湖南农业大学学报（自然科学版）》2022年第6期691-698,共8页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(31720103915);湖南省自然科学基金项目(2020JJ4040);湖南省科学技术厅湖湘高层次人才聚集工程项目(2019RS1052);湖南省教育厅研究生科研创新项目(CX20200665)。

摘　　要：采集湖南省6个柑橘产区的呈现典型溃疡病症状的柑橘叶片、枝条或果实样品26份,分离溃疡病菌株,采用柑橘溃疡病特异性引物鉴定、细菌保守的16S rDNA序列分析及重复序列PCR基因指纹分析(Rep-PCR)方法,分析具有地域代表性的12个菌株间基因组多态性。结果显示,12个菌株在BOX-PCR和ERIC-PCR中扩增出相同数量的多态性条带,无法通过Rep-PCR对菌株进行区分;串联重复位点多态性分析表明,CCB-2、CCB-3和CCB-10这3个菌株在GT1位点上出现片段大小多态性;通过活体注射接种方式,将12个溃疡病菌株接种至冰糖橙叶片,结果 CCB-2和CCB-4菌株表现出较稳定致病力,综合病情指数达到75%以上,表明CCB-2相比其他11个菌株可能在基因组序列水平存在多态性,且影响菌株的致病力水平。Twenty-six citrus leaf, branch or fruit samples showing typical symptoms caused by Xanthomonas citri subsp.citri(Xcc) were collected from six citrus producing areas in Hunan Province, and the citrus canker pathogen strains were isolated. Genomic polymorphisms among 12 geographically representative pathogen strains were analyzed by bacterial conserved 16S r DNA sequence analysis and repetitive sequence PCR gene fingerprinting(Rep-PCR) methods using citrus canker-specific primer. The results showed that the 12 strains showed the same number of polymorphic bands in BOX-PCR and ERIC-PCR and could not be distinguished by Rep-PCR, while tandem repeat polymorphism analysis showed that 3 strains, strain CCB-2, strain CCB-3 and strain CCB-10, showed fragment size polymorphism at the GT1locus. Twelve strains of citrus canker were inoculated by in vivo injection into the leaves of ‘Bingtang’ sweet orange and two strains, strain CCB-2 and strain CCB-4, showed stable pathogenicity with a combined disease index of over 75%,indicating that strain CCB-2 may have polymorphism at the genomic sequence level compared to the other 11 strains and affect the pathogenicity level of the strain.

关 键 词：柑橘 柑橘溃疡病 多态性 致病力 湖南 

分 类 号：S436.661[农业科学—农业昆虫与害虫防治]